1.Expression and purification of nucleocapsid protein of MERS coronavirus in E.coli
Yangbo FU ; Yong HU ; Chengcheng HUANG ; Yuanyuan BAI ; Lihong QIU ; Cheng CAO ; Ting GAO
Military Medical Sciences 2015;39(12):919-922
Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.
2.Construction and application of network education platform of ophthalmology
Guiqiu ZHAO ; Chengcheng ZHU ; Liting HU ; Qiang XU ; Nan JIANG ; Sheng QIU
Chinese Journal of Medical Education Research 2014;(8):861-864
The network teaching platform of ophthalmology of Qingdao University , as the basis construction of the national key discipline , forms a perfect and complete set of teaching system with the aid of part of course information, part of network teaching resources and part of answer and interaction. The network teaching platform focuses on the construction of network teaching resources and answer and interaction. In the college teaching of ophthalmology, through building clinical teaching resource database and discussing on network platform, we carry out interactive and discussion-based teaching, and students can prepare before class and discuss after class. The application of network teaching platform of ophthal-mology in classroom teaching and teaching feedback can improve the teaching methods , deepen the teach-ing content, implement the sharing of teaching resources, and lay a solid foundation for ophthalmology teaching reform.
3.Non-T cell depletion haploidentical hematopoietic stem-cell transplantation for T lymphoblastic lymphoma
Xuefeng HE ; Depei WU ; Aining SUN ; Xiang ZHANG ; Huiying QIU ; Yue HAN ; Xiaowen TANG ; Chengcheng FU ; Zhengming JIN
Journal of Leukemia & Lymphoma 2011;20(8):459-463
Objective To explore the efficacy of non-T cell depletion haploidentical hematopoietic stem-cell transplantation for T lymphoblastic lymphoma (T-LL). Methods 3 T-LL patients achieving complete remission received haploidentical bone marrow stem cell transplantation with granulocyte-colony-stimulating factor (G-CSF) mobilized bone marrow grafts from related donor without T-cell depletion. Two of them received a myeloablative conditioning regimen consisting of high-doses of cyclophosphamide and cytarabine with total body irradiation, whereas the other was preconditioned with busulfan, cyclophosphamide and cytarabine. All patients received strengthened phophylaxis regimen including rabbit anti-thymocyte globulin against acute graft-versus-host disease. Results All patients had rapid hematopoietic engraftment with the median time for neutrophil and platelet recovery being 12 days and 13 days, respectively. They are still alive without relapse at a median follow-up of 24 months (range: 9-75 months). Conclusion Treatment related toxicity can be acceptable in non-T cell depletion haploidenfical hematopoietic stem-cell transplantation for T-LL and the patients may achieve long term survival.
4.Preliminary analysis on graft failure after non-T-cell depleted haploidentical hematopoietic stem cell transplantation
Bin GU ; Guanghua CHEN ; Xiao MA ; Chengcheng FU ; Yue HAN ; Xiaowen TANG ; Zhengming JIN ; Miao MIAO ; Huiying QIU ; Aining SUN ; Depei WU
The Journal of Practical Medicine 2016;32(20):3401-3404
Objective To summarize the clinical features of graft failure (GF)after non-T-cell depleted haploidentical hematopoietic stem cell transplantation (Haplo-HCT), and to investigate the causes and treatment. Methods A retrospective analysis was carried out on 174 patientswho accepted the non-T-cell depleted Haplo-HCT from Jan 2012 to Dec 2013. The patients′ donor specific anti human leukocyte antigen antibodies (DSA) from the peripheral blood serum were detected and those DSA positive patients were treated by immunoglobulin or plasma exchange before transplatation. Results A total of three patients with acute myeloid leukemia got GF, the incidence rate was 1.72%. The patient with primary GF was given a secondHaplo-HCT, but did not get implanted with leukemia remission and three lineages persistently low , he was died of pulmonary infection eight monthes after the second transplant. One of the secondary GF patients was given peripheral blood mononuclear cells(PBMNCs) mobilized by granulocyte colony stimulating factor (G-CSF) from the donor, and got full donor chimerism on day 16 after infusion. The disease-free survival has been for 18 months. The other case was found that DSA was positive, the mean fluorescence intensity (MFI) value was 15000, then Rituximab and PBMNCs mobilized by G-CSF were administrated successively. On day 14 after infusion the partient got full donor chimerism , and MFI turned negative. The patient has been disease-free survival for 41 months. Conclusion Graft failure is a rare but fatal complication after non-T-cell depletedHaplo-HCT, Rituximab followed by PBMNCs are effective measures for DSA related GF, as were worthy of further study.
5. Comparison of the efficacy of decitabine combined with micro-transplantation or priming regimen as consolidation treatment for older patients with acute myeloid leukemia
Weiyang LI ; Yufeng FENG ; Xiao MA ; Huiying QIU ; Chengcheng FU ; Xiaowen TANG ; Yue HAN ; Deipei WU ; Aining SUN
Chinese Journal of Hematology 2018;39(4):305-309
Objective:
To probe the feasibility of decitabine (DAC) combined with micro-transplantation as consolidation treatment for older patients with acute myeloid leukemia (AML).
Methods:
Between November 2012 and September 2015, 37 consecutive patients with AML ≥60 years of age were analyzed. Of them, 19 patients received consolidation therapy with DAC followed by micro-transplantation (microtransplant group). Another 18 ones (chemo group) were treated with DAC plus priming regimen as consolidation chemotherapy in the same period.
Results:
There were no significant differences in terms of age, WBC count, and disease status of onset between the microtransplant and chemo groups (
6. Comparison of efficacy of first-line administration of generic dasatinib or imatinib in patients with Philadelphia chromosome positive acute lymphoblastic leukemia treated by hematopoietic stem cell transplantation
Fei YANG ; Wenzhi CAI ; Xiaowen HUANG ; Shengli XUE ; Chengcheng FU ; Xiaowen TANG ; Aining SUN ; Depei WU ; Huiying QIU
Chinese Journal of Hematology 2018;39(8):661-667
Objective:
To investigate the efficacy of first-line administration of generic dasatinib or first-generation TKI (imatinib) in patients with Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ ALL) treated by hematopoietic stem cell transplantation (HSCT).
Methods:
Clinical features and prognoses of 63 newly diagnosed Ph+ ALL patients from Jan 2014 to June 2017 treated by HSCT combined with first-line administration of generic dasatinib or imatinib were retrospective analyzed.
Results:
Of 63 Ph+ ALL patients, 31 cases were administered generic dasatinib, and the other 32 ones imatinib. Complete remission (CR) rates at the fourth week of induction therapy in generic dasatinib and imatinib groups were 96.8% and 93.8% (
7.Value of ultrasound-based radiomics combined with clinical parameters in predicting lymph node metastasis of endometrial cancer
Lisha QIN ; Chengcheng QIU ; Zhiping TANG ; Lijuan LI ; Tingting LIU ; Yan MA
Chinese Journal of Ultrasonography 2022;31(4):332-337
Objective:To explore the value of ultrasound-based radiomics combined with clinical parameters in predicting lymph node metastasis of endometrial cancer.Methods:The clinical data and preoperative ultrasound images of 255 patients with endometrial cancer diagnosed after operation in the First Affiliated Hospital of Guangxi Medical University from January 2017 to February 2021 were analyzed retrospectively. The boundary of the tumor, that is, the region of interest(ROI), was determined manually by ITK-SNAP software, and the features were extracted from it. Through random sampling, the data were divided into training group used to build the model and verification group used to evaluate the reliability of the model at the proportion of 7∶3.In the training group, the spearman correlation coefficient was used as the threshold of 0.95 to remove the features with high correlation. The importance of the features were ranked by the decision tree model, the features of the top 6% importance were selected to establish a logical regression model, and then calculated the Radiomics score of each patient. On the basis of logical regression analysis, a comprehensive prediction model was established, the line diagram was drawn, and the prediction effect was evaluated by the analysis of the area under the subject working curve (AUC), calibration curve and the decision curve.Results:After dimensionality reduction and logical regression analysis, 20 features and 2 clinical parameters (CA125, CA153) were obtained to build the model. The AUC of training group and verification group in radiomics model and comprehensive prediction model were 0.81, 0.78 and 0.89, 0.85, respectively. The decision curve analysis showed that both of them had good clinical utility.Conclusions:The ultrasound-based radiomics model and the comprehensive model combined with relevant clinical parameters are effective in predicting lymph node metastasis of endometrial cancer.
8. Comparison of haploidentical hematopoietic stem cell transplantation and matched-sibling donor transplantation for the treatment of paroxysmal nocturnal hemoglobinuria
Limin LIU ; Huifen ZHOU ; Qingyuan WANG ; Huiying QIU ; Xiaowen TANG ; Yue HAN ; Chengcheng FU ; Zhengming JIN ; Suning CHEN ; Aining SUN ; Miao MIAO ; Depei WU
Chinese Journal of Hematology 2019;40(4):306-311
Objective:
To compare the outcomes between haploidentical donor hematopoietic stem cell transplantation (haplo-HSCT) and matched-sibling donor transplantation (MSD-HSCT) for paroxysmal nocturnal hemoglobinuria (PNH) .
Methods:
The clinical data of 40 PNH patients received HSCT (haplo-HSCT=25, MSD-HSCT=15) from July 2007 to May 2018 were analyzed retrospectively to compare the outcomes between haplo-HSCT and MSD-HSCT groups.
Results:
There were no differences in terms of gender, age, patients of PNH-AA and median time from diagnosis to transplantation between the 2 groups (
9. Outcomes of allogeneic hematopoietic stem cell transplantation in patients with paroxysmal nocturnal hemoglobinuria compared to paroxysmal nocturnal hemoglobinuria-aplastic anemia syndrome
Limin LIU ; Huifen ZHOU ; Qingyuan WANG ; Huiying QIU ; Xiaowen TANG ; Yue HAN ; Chengcheng FU ; Zhengming JIN ; Suning CHEN ; Aining SUN ; Miao MIAO ; Depei WU
Chinese Journal of Hematology 2019;40(6):472-476
Objective:
To compare the outcomes of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for paroxysmal nocturnal hemoglobinuria (PNH) with paroxysmal nocturnal hemoglobinuria-aplastic anemia (PNH-AA) syndrome.
Methods:
The outcomes of 46 patients who received allo-HSCT (16 PNH patients, 30 PNH-AA patients) from July 10, 2007 to June 2, 2018 were analyzed retrospectively. The conditioning regimen was busulfan, cyclophosphoramide, and ATG in haploidentical donors and unrelated donors. Patients with matched sibling donors were treated with the fludarabine, cyclophosphamide, and ATG regimen.
Results:
There were no differences of baseline data between the 2 groups except gender distribution and the numbers of haploidentical donor transplantation. The median values of absolute nucleated cell counts were 10.58 (3.83-13.83) ×108/kg in the PNH group and 10.81 (3.96-33.40) ×108/kg in the PNH-AA group (
10. Clinical significance of JAK2、CALR and MPL gene mutations in 1 648 Philadelphia chromosome negative myeloproliferative neoplasms patients from a single center
Mengyun LI ; Hongying CHAO ; Aining SUN ; Huiying QIU ; Zhengming JIN ; Xiaowen TANG ; Yue HAN ; Chengcheng FU ; Suning CHEN ; Depei WU
Chinese Journal of Hematology 2017;38(4):295-300
Objective:
To explore the prevalences of JAK2, CALR and MPL gene mutations and the mutation types in patients with Philadelphia chromosome negative myeloproliferative neoplasms (MPNs) , and to compare their clinical characteristics of different mutation types with each other and mutation negative group.
Methods:
The mutations of JAK2 V617F, JAK2 gene at exon 12, CALR gene at exon 9 and MPL gene at exon 10 in 1 648 Ph negative MPNs patients were detected by direct sequencing.
Results:
① The JAK2V617F mutation was found in 471 (92.7%) of 508 PV patients, 819 (78.1%) of 1 049 ET patients and 74 (81.3%) of 91 PMF patients respectively, with the total mutation rate as 82.8% (1 364/1 648) . The JAK2 exon12 mutation was found in 9 (1.7%) of 508 PV patients, none was found in ET or PMF patients, with the total mutation rate as 0.5% (9/1 648) . The CALR mutation was found in 132 (12.6%) of 1 049 ET patients and 11 (12.1%) of 91 PMF patients respectively, with the total mutation rate as 8.7% (143/1 648) ; the MPL mutation was found in 9 (0.9%) of 1 049 ET patients and 1 (1.1%) of 91 PMF patients respectively, with the total mutation rate as 0.6% (10/1 648) . The co-occurrence of any two types of driver gene mutations was not detected by direct sequencing. ②The median onset age of patients with JAK2V617F[61 (15-95) y] was significant higher than of with JAK2 exon12 mutation[49 (33-62) y] or without mutations[42 (3-78) y] (